Main Article Content
Jatropha curcas is one of the potential biodiesel production plant. However, one serious problem is that the plant produces phorbol esters (PEs), a cacinogen. Geranylgeranyl diphosphate synthase (GGPPS) is a key enzyme in the phorbol esters biosynthesis. To study the role of GGPPS gene, the partial GGPPS gene was cloned by using RT-PCR from total RNA extracted from jatropha seed cv. Korat and with the primer specific to GGPPS gene. The DNA fragment size of 592 bp was amplified. The sequence analysis indicated that the obtained cDNA had very high similarity (99 %) to GGPPS gene of J. curcas. We named this DNA fragment as JcGGPPS. The antisense JcGGPPS gene was introduced into tobacco by Agrobacterium tumefacients strain EHA-105. The four transgenic tobacco plants were produced. The analysis of GGPPS gene expression by real-time PCR showed that the lowest expression level of endogenous GGPPS gene in transformed plants was about 3 folds lower than that of the control plant. The phorbol esters contents of transformed tobacco plants were determined by high performance liquid chromatography (HPLC). It was found that the PEs content of the transformed tobacco was also 2.5 folds lower than that of the control. It was indicated that the antisense JcGGPPS gene was an effective down-regulating in the expression of endogenous GGPPS gene and reducing the PEs content of tobacco.
 Heller, J., 1996, Physic Nut – Jatropha curcas L. – Promoting the Conservation and Use of Underutilized and Neglected Crops, Doctoral Dissertation, Institute of Plant Genetic and Crop Plant Research, Gatersleben, Germany & International Plant Genetic Resource Institute, Rome.
 Devappa, R.K., Makkar, H.P.S. and Becker, K., 2010a, Jatropha toxicity: A review, J. Toxicol. Environ. Health B Crit. Rev. 13: 476-507.
 Devappa, R.K., Makkar, H.P.S. and Becker, K., 2010b, Optimization of conditions for the extraction of phorbol esters from Jatropha oil, Biomass Bioenerg. 34: 1125-1133.
 Yunping, B., Ha, B.T.N., Eunice, Y., Chueng, L.L. and Yan, H., 2012, Light induced degradation of phorbol esters, Ecotoxicol. Environ. Saf. 84: 268-273.
 Kaewsuriwong, S., Comwien, J., Roytrakul, S. and Chulalaksananukul, W., 2013, Proteomic techniques for analysis of toxin biosynthetic pathways in Jatropha curcas, Thai J. Genet. 6: 115-127. (in Thai)
 Ahmed, W.A. and Salimon, J., 2009, Phorbol esters as constituents of tropical Jatropha curcas oil. Eur. J. Sci. Res. 31: 429-436.
 Devappa, R.K., Makkar, H.P.S. and Becker, K., 2011, Jatropha diterpenes: A review, J. Am. Oil Chem. Soc. 88: 301-322.
 Lin, J., Jin, Y., Zhou, X. and Wang, J.Y., 2010, Molecular cloning and functional analysis of the gene encoding geranyl geranyl diphosphate synthase from Jatropha curcas, Afr. J. Biotechnol. 9: 3342-3351.
 Laksana, C. and Chanprame, S., 2015, A simple and rapid method for RNA extraction from young and mature leaves of oil palm (Elaeis guineensis Jacq.), J.
ISSAAS. 21: 96-106.
 Singlo, W., 2012, Transformation of Glyphosate Resistance Gene into Physic Nut (Jatropha curcas L.), Master Thesis, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom, 87 p. (in Thai)
 Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W., 1987, GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants, EMBO J. 6: 3901-3907.
 Amkul, K., Laosatit, K., Somta, P., Shim, S., Lee, SH., Tanya P. and Srinives. P., 2017, Mapping of QTLs for seed phorbol esters, a toxic chemical in Jatropha curcas (L.), Genes 8: 205.
 Hinjan, S., 2010, Transformation of Antisense LIM Gene into Camaldulensis Dehnh, Doctoral Dissertation, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom, 120 p. (in Thai)
 Shan, W., Zhao, C., Fan, J., Cong, H., Liang, S. and Yu, X., 2012, Antisense suppression of alcohol acetyltransferase gene in ripening melon fruit alters volatile composition, Sci Hort. 139: 96-101.
 Jassbi, A.M., Gsae, K., Hettenhausen, C., Schmidt, A. and Baldwin, I., 2008, Silencing geranylgeranyl diphosphate synthase in Nicotiana attenuata dramatically impairs resistance to tobacco hornworm, Plant Physiol. 146: 974-986.
 Haas, W., Sterk, H. and Mittelbach, M., 2002, Novel 12-deoxy-16-hydroxyphorbol diesters isolated from the seed oil of Jatropha curcas, J. Nat. Prod. 65: 1434-1440.
 Martinez-Herrera, J., Siddhuraju, P., Francis, G., Dávila-Ortíz, G. and Becker, K., 2006, Chemical composition, toxic/ antimetabolic constituents, and effects of different treatments on their levels, in four provenances of Jatropha curcas L. from Mexico, Food Chem. 96: 80-89.
 Katoh, A., Ohki, H., Inai, K. and Hashimoto, T., 2005, Molecular regulation of nicotine biosynthesis, Plant Biotechnol. J. 22: 389-392.
 Okada, K., Sato, T., Nakagawa, T., Kawamukai, M. and Kamiya, Y., 2000, Five geranylgeranyl diphosphate synthase expressed in different organs are localized into three subcellular compartments in Arabidopsis, Plant Physiol. 122: 1045-1056.